EC Number | Cloned (Comment) | Organism |
---|---|---|
3.4.21.88 | gene lexA, recombinant expression of His-tagged wild-type LexA and mutant enzymes S160A, K197A, and G126D, the isolated C-terminal segment and the N-domain in Escherichia coli strain BL21(DE3) pLysS | Mycobacterium tuberculosis |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
3.4.21.88 | purified recombinant His-tagged wild-type LexA and mutant enzymes S160A, K197A, and G126D, the isolated C-terminal segment and the N-domain, wild-type LexA diffraction-quality crystals grow by vapour diffusion method in about three months from a drop consisting of 0.002 ml of 6 mg/ml protein solution and 0.002 ml of 100 mM bis-Tris, pH 6.5 or pH 8.5, 100 mM NaCl or 200 mM MgCl2, 5% glycerol, 25% PEG 3350 (form I and form II, respectively). The microbatch-under-oil method is used for crystallization of the C-domain by mixing of 0.002 ml of 10 mg/ml protein solution with 0.002 ml 100 mM Tris-HCl, pH 7.5, 25% glycerol, and 40% v/v pentaerythritol ethoxylate (15/4 EO/OH). Diffraction-quality crystals of the S160A mutant are obtained by microbatch-under-oil method and mixing of 0.002 ml 10 mg/ml protein and 0.002 ml 100 mM bis-Tris, pH 6.5, 200 mM MgCl2, and 25% w/v PEG 3350 (form III), and of mutant K197A by mixing of 0.002 ml 10 mg/ml protein solution with 0.002 ml crystallization solution containing 100 mM HEPES, pH 7.5, 20 mM MgCl2, and 22% w/v polyacrylic acid sodium salt (form IV), while crystals of the G126D mutant are grown from a solution consisting of 0.002 ml of 10 mg/ml protein with 0.002 ml of 100 mM Tris-HCl, pH 8.5, 100 mM sodium acetate trihydrate, and 30% w/v PEG 4000 (form V), mutant crystals grow about 2 months, X-ray diffraction structure determination and analysis at 1.48-2.25 A resolution | Mycobacterium tuberculosis |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
3.4.21.88 | G126D | site-directed mutagenesis, cleavage site mutant | Mycobacterium tuberculosis |
3.4.21.88 | K197A | site-directed mutagenesis, active site mutant | Mycobacterium tuberculosis |
3.4.21.88 | S160A | site-directed mutagenesis, active site mutant | Mycobacterium tuberculosis |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
3.4.21.88 | NaCl | variation of the sodium chloride concentration from 25 mM to 1.5 M do not show any significant change in the autoproteolysis of MtLexA | Mycobacterium tuberculosis |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
3.4.21.88 | Mycobacterium tuberculosis | P9WHR7 | - |
- |
3.4.21.88 | Mycobacterium tuberculosis ATCC 25618 | P9WHR7 | - |
- |
3.4.21.88 | Mycobacterium tuberculosis H37Rv | P9WHR7 | - |
- |
EC Number | Posttranslational Modification | Comment | Organism |
---|---|---|---|
3.4.21.88 | proteolytic modification | the autocatalytic cleavage of MtLexA and the mutant proteins is analyzed, effects of pH and temperature, the enzyme is autocatalytically active at pH 9.5-11.0 and at up to 55°C | Mycobacterium tuberculosis |
EC Number | Purification (Comment) | Organism |
---|---|---|
3.4.21.88 | recombinant His-tagged wild-type and mutant enzymes, as well as enzyme fragments from Escherichia coli strain BL21(DE3) pLysS by nickel affinity chromatography and gel filtration, to over 95% purity | Mycobacterium tuberculosis |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
3.4.21.88 | additional information | qualitative analysis of DNA-binding activities of MtLexA and its mutants, MtLexA binds to dsDNA with the consensus sequence GAAC-N4-GTTT/C. Modeling of the MtLexA-DNA complex, overview. The length of the flanking sequences of bound DNA does not affect DNA binding by LexA. Analysis of autocatalytic cleavage of MtLexA and enzyme mutants | Mycobacterium tuberculosis | ? | - |
? | |
3.4.21.88 | additional information | qualitative analysis of DNA-binding activities of MtLexA and its mutants, MtLexA binds to dsDNA with the consensus sequence GAAC-N4-GTTT/C. Modeling of the MtLexA-DNA complex, overview. The length of the flanking sequences of bound DNA does not affect DNA binding by LexA. Analysis of autocatalytic cleavage of MtLexA and enzyme mutants | Mycobacterium tuberculosis H37Rv | ? | - |
? | |
3.4.21.88 | additional information | qualitative analysis of DNA-binding activities of MtLexA and its mutants, MtLexA binds to dsDNA with the consensus sequence GAAC-N4-GTTT/C. Modeling of the MtLexA-DNA complex, overview. The length of the flanking sequences of bound DNA does not affect DNA binding by LexA. Analysis of autocatalytic cleavage of MtLexA and enzyme mutants | Mycobacterium tuberculosis ATCC 25618 | ? | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
3.4.21.88 | homodimer | 2 * 12000, C-terminal catalytic core, SDS-PAGE | Mycobacterium tuberculosis |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
3.4.21.88 | MtLexA | - |
Mycobacterium tuberculosis |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.21.88 | 37 | - |
assay at | Mycobacterium tuberculosis |
EC Number | Temperature Minimum [°C] | Temperature Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.21.88 | 37 | 55 | self-cleavage activity at | Mycobacterium tuberculosis |
EC Number | Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|---|
3.4.21.88 | 55 | - |
above, melting temperature of wild-type and mutant enzymes | Mycobacterium tuberculosis |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
3.4.21.88 | 9.5 | 11 | self-cleavage activity at | Mycobacterium tuberculosis |
EC Number | General Information | Comment | Organism |
---|---|---|---|
3.4.21.88 | additional information | a complex of Mycobacterium tuberculosis LexA and the cognate SOS box is modeled in which the mutual orientation of the two N-terminal domains differs from that in the Escherichia coli LexA-DNA complex, complex structure analysis, overview | Mycobacterium tuberculosis |
3.4.21.88 | physiological function | LexA is a critical protein involved in the bacterial SOS response, which consists of the coordinated activation of a network of genes required for DNA repair and mutagenesis in response to DNA damage. The clinical relevance of the SOS response in bacteria can be attributed not only to its involvement in virulence and mutagenesis, but also in the spread of antibiotic resistance | Mycobacterium tuberculosis |